VEGF-C反义寡核苷酸对卵巢癌细胞株SKOV3侵袭转移能力的影响

目的]探讨VEGF—C反义寡核苷酸（VEGF—CASODN）对人卵巢癌细胞SKOV3侵袭转移能力的影响。方法]以VEGF—C基因编码区为靶点合成反义寡核苷酸，脂质体介导转染。MTT法检测卵巢癌细胞与细胞外基质Matrigel的黏附能力；体外侵袭实验检测卵巢癌细胞侵袭能力；Western blot检测细胞VEGF—C、MMP-2、E—cadherin蛋白表达。结果]黏附实验结果显示VEGF—C ASODN转染后，在30min、60min、90min、120min各时间段与空白对照组、LIP组及SODN组相比，细胞黏附率都有明显下降（P〈0．01）：侵袭实验结果显示VEGF—CASODN转染48h后浸润细胞数目（62．5±8．71与空白对照组、LIF组和SODN组（分别为127.2±13．6、125．8±14．1、119．3±11．21相比明显减少（P〈0．01）；Western blot结果显示VEGF—C ASODN转染后细胞中VEGF—C、MMP-2蛋白表达与空白对照组、LIF组和SODN组相比明显下降，而E—cadherin蛋白表达明显增高（P〈0．01）。结论]VEGF—C ASODN能明显抑制卵巢癌细胞体外实验中的侵袭转移能力，可望为卵巢癌提供一种有效的辅助治疗方法。

Effect of VEGF-C Antisense Oligodeoxvnucleotide(ASODN) on Invasion and Metastasis of Ovarian Cancer Cells SKOV3

Purpose ] To investigate the effect of VEGF-C antisense oligodeoxynucleotide（ASODN） on the invasion and metastasis of human ovarian cancer cell line SKOV3. Methodsl The ASODN sequence targeting the coding region of VEGF-C was synthesized and transfected into ovarian cancer cells by liposomes.The adhesion of SKOV3 cells to artifical basement membrane Matrigel was measured by MTT assay. The invasion was evaluated by transwell chambers attached with polycarbonate filters and reconstituted basement membrane（Matrigel）. The expression of VEGF-C, MMP-2 and E-cadherin protein was measured by Western blot assay. Result] The result of adhesion experiment demonstrated that VEGF-C ASODN markedly suppressed the adhesion ability of SKOV3 cells to Matrigel at 30min, 60min, 90min and 120min compared with the control group and the lipofectin （LIP） group and the sense oligodeoxynucleotide （SODN） group （P〈0.01）. The result of invasion experiment demonstrated the invasive cell number in VEGF-C ASODN group （62.5±8.7） decreased significantly than that in the control group, the lipofectin group and SODN group respectively （127.2±13.6, 125.8±14.1 and 119.3±11.2, P〈0.01）. The result of Western blot showed the expression of VEGF-C and MMP-2 protein in VEGF-C ASODN group was remarkably downregulated and E-cadherin protein up-regulated compared with the control group, the lipofectin group and SODN group （P〈0.01）. Conclusionl VEGF-C ASODN can inhibit invasion and metastasis of SKOV3 cells remarkably, which might become an effective adjuvant therapy for ovarian cancer in the future.